The efficacy and safety of paravertebral block for postoperative analgesia in renal surgery: A systematic review and meta-analysis of randomized controlled trials.

Background: Paravertebral block (PVB) has been widely used in postoperative analgesia, especially in thoracic and breast surgery. However, the efficacy and safety of PVB for analgesia after renal surgery remains uncertain. Therefore, this study aimed to determine the postoperative analgesic efficacy and safety of PVB in renal surgery. Methods: PubMed, Web of Science, Embase, and the Cochrane Library databases were systematically searched up to December 20, 2021. All randomized controlled trials (RCTs) evaluating the postoperative analgesic efficacy of PVB in renal surgery were collected. The meta-analysis was performed using RevMan 5.4 and Stata/MP 14.0 software. Results: A total of 16 RCTs involving 907 patients were included in the meta-analysis. Ten studies investigated patients under percutaneous nephrolithotomy (PCNL), and six studies were done for patients under other renal surgery (nephrectomy or pyeloplasty). Compared with control groups (no block, sham block, or other nerve blocks), meta-analysis showed that PVB reduced 24-hour postoperative opioid consumption significantly (SMD = -0.99, 95%CI: -1.60-0.38, p = 0.001, I 2 = 92%) and reduced pain scores at various time points within 24 h at rest and 1 h, 4 h, and 24 h at movement after renal surgery, furthermore, PVB prolonged the time to first postoperative analgesic requirement (SMD = 2.16, 95%CI: 0.94-3.39, p = 0.005, I 2 = 96%) and reduced the incidence of postoperative additional analgesia (OR = 0.14, 95%CI: 0.06∼0.33, p < 0.00001, I 2 = 50%). Subgroup analysis revealed that the postoperative analgesia effect of PVB was more significant in PCNL, and the use of bupivacaine for PVB seemed to have a better performance. Besides, there was no difference in the incidence of postoperative nausea, vomiting, and itching between PVB and control groups. Conclusion: This study indicates that PVB may provide effective postoperative analgesia in patients under renal surgery, especially PCNL patients. Moreover, PVB is a safe analgesic method without significant analgesia-related complications.

Expression of CHL1 in Clear Cell Renal Cell Carcinoma and its Association With Prognosis.

As a member of the L1 family of neural cell molecules, close homologue of L1 (CHL1) has been proved to be downregulated in several human cancers. In the present study, we aimed to assess the expression and prognostic value of CHL1 in clear cell renal cell carcinoma (CCRCC). Immunohistochemistry was performed to detect the expression of CHL1 in tissue microarray chips. Then we compared specific clinicopathologic features in patients with different CHL1 expressions. The correlation between CHL1 expression and overall survival (OS) was evaluated by the Kaplan-Meier method and Cox regression analysis. We found that the expression of CHL1 was significantly lower in CCRCC tissues compared with adjacent normal tissues, which was correlated with TNM stage (P<0.001), Fuhrman grade (P=0.006), and LVI (P=0.004). The Kaplan-Meier survival analysis indicated that CCRCC patients with low CHL1 expression had a poorer OS rate than those with high CHL1 expression (P<0.001). Univariate and multivariate Cox regression analyses suggested that CHL1 was an independent and unfavorable prognostic factor for the OS rate of CCRCC patients. Collectively, low expression of CHL1 might predict poor OS rate of CCRCC.

Association of Paraoxonase 1 Gene Polymorphisms with the Risk of Breast Fibroadenoma and Breast Cancer in the Females of Guangxi, China.

Fibroadenoma of the breast is a common cause of a benign breast lump in premenopausal women. The consensus view is that women with fibroadenomas are not at significantly increased risk of developing breast cancer. The objective of this research was to explore the association of PON1 rs662 and rs705382 with the risk of breast fibroadenoma (BF) and breast cancer (BC) in the females of Guangxi in southern China. The PON1 rs662 and rs705382 single-nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 55 BF patients, 80 BC patients, and 98 healthy controls. Significant associations with FB and BC risks were observed for the rs662 SNP. Diagnosis is based on the combination of clinical examination, imaging, and nonsurgical tissue biopsy (the triple test) [21]. In haplotype analyses, the haplotype GA increases the risk and haploid GG decreases the risk in BF and BC. Our research indicated that the PON1 rs662 SNP might be a risk factor for BF and BC. The results of this research indicated that the locus of rs662 in PON1 is relevant to risk of developing BF and BC in the females of Guangxi. Further prospective studies are needed to support our conclusions.

[Risk factors for postoperative long-term hematuria in patients with benign prostatic hyperplasia].

OBJECTIVE: To explore the risk factors for long-term hematuria after operation in BPH patients. METHODS: We retrospectively analyzed the clinical data on 646 cases of BPH treated by transurethral surgery in Liyang People's Hospital from January 2015 to August 2020. According to the incidence of hematuria at 3 months or longer after surgery, we divided the patients into a hematuria and a non-hematuria group, recorded the related factors, and investigated the independent risk factors for long-term hematuria by univariate and multivariate analyses. RESULTS: Of the 646 BPH patients, 48 were found with and 598 without hematuria after transurethral surgery. Univariate analysis showed that hypertension, diabetes mellitus, residual prostate gland, urinary tract infection, bladder neck contracture, prostate cancer, urethral calculus, urethral stricture, excessive activity and constipation were the influencing factors (P < 0.05), while multivariate logistic regression analysis revealed that hypertension (P < 0.001), diabetes mellitus (P = 0.007), residual prostate gland (P = 0.013), urinary tract infection (P < 0.001), bladder neck contracture (P = 0.032), urethral calculus (P = 0.033) and urethral stricture (P = 0.001) were independent risk factors for long-term hematuria in the BPH patients after surgery. CONCLUSION: Complicated hypertension, diabetes mellitus, residual prostate gland, urinary tract infection, bladder neck contracture, urethral calculus and urethral stricture are independent risk factors for long-term hematuria in BPH patients after transurethral surgery.

Capn4 contributes to tumor invasion and metastasis in clear cell renal cell carcinoma cells via modulating talin-focal adhesion kinase signaling pathway.

Calpain small subunit 1 (Capn4) has been shown to correlate with the metastasis/invasion of clear cell renal cell carcinoma (ccRCC). This study aimed to further elucidate the molecular mechanisms underlying Capn4-mediated ccRCC progression. The mRNA expression levels in ccRCC cells were measured by quantitative real-time PCR. The effects of Capn4 on cell adhesion, invasion, and migration were examined by cell adhesion assay, cell invasion assay, and wound-healing assay, respectively. The protein levels were detected by western blot analysis. The effect of Capn4 on cancer metastasis in vivo was assessed in a nude mice xenograft model. It was found that Capn4 was up-regulated in the ccRCC cells, and Capn4 overexpression suppressed cell adhesion activity and increased cell invasion and migration in 786-O cells, while Capn4 silencing increased cell adhesion activity and impaired the invasion and migration ability of Caki-1 cells. Capn4 overexpression also increased the protein level of cleaved talin in 786-O cells, while Capn4 silencing decreased the protein level of cleaved talin in Caki-1 cells. The focal adhesion kinase (FAK)/AKT/MAPK signaling was activated by Capn4 overexpression in 786-O cells, and was inhibited by Capn4 down-regulation in Caki-1 cells. Capn4 overexpression increased the protein levels of matrix metalloproteinase 2 (MMP-2), vimentin, N-cadherin, and down-regulated E-cadherin in 786-O cells, while Capn4 silencing decreased the protein levels of MMP-2, vimentin, N-cadherin, and up-regulated E-cadherin in Caki-1 cells. Capn4 also promoted cancer metastasis in the in vivo nude mice xenograft model. Our results implicate the functional role of Capn4 in ccRCC invasion and migration, which may contribute to cancer metastasis in ccRCC.

Fruit and vegetable intake and breast cancer prognosis: a meta-analysis of prospective cohort studies.

The effect of fruit and vegetable intake on breast cancer prognosis is controversial. Thus, a meta-analysis was carried out to explore their associations. A comprehensive search was conducted in PubMed, Web of Science, OVID, ProQuest and Chinese databases from inception to April 2016. The summary hazard ratios (HR) and 95 % CI were estimated using a random effects model if substantial heterogeneity existed and using a fixed effects model if not. Subgroup analyses and sensitivity analyses were also performed. In total, twelve studies comprising 41 185 participants were included in the meta-analysis. Comparing the highest with the lowest, the summary HR for all-cause mortality were 1·01 (95 % CI 0·72, 1·42) for fruits and vegetables combined, 0·96 (95 % CI 0·83, 1·12) for total vegetable intake, 0·99 (95 % CI 0·89, 1·11) for cruciferous vegetable intake and 0·88 (95 % CI 0·74, 1·05) for fruit intake; those for breast cancer-specific mortality were 1·05 (95 % CI 0·77, 1·43) for total vegetable intake and 0·94 (95 % CI 0·69, 1·26) for fruit intake; and those for breast cancer recurrence were 0·89 (95 % CI 0·53, 1·50) for total vegetable intake and 0·98 (95 % CI 0·76, 1·26) for cruciferous vegetable intake. This meta-analysis found no significant associations between fruit and vegetable intake and breast cancer prognosis.

Serum betaine but not choline is inversely associated with breast cancer risk: a case-control study in China.

PURPOSE: Choline and betaine are important for DNA methylation and synthesis, and may affect tumor carcinogenesis. To our knowledge, no previous study has examined the association between serum choline and betaine and breast cancer risk. This study aimed to examine whether serum choline and betaine were inversely associated with breast cancer risk among Chinese women. METHODS: This hospital-based case-control study consecutively recruited 510 breast cancer cases and 518 frequency-matched (age and residence) controls, and blood samples were available for 500 cases and 500 controls. Serum choline and betaine were assayed by high-performance liquid chromatography-tandem mass spectrometry. Multiple unconditional logistic regression was used to estimate odds ratios (ORs) and 95 % confidence intervals (CIs). RESULTS: An inverse association with breast cancer risk was observed for serum betaine (fourth vs first quartile adjusted OR 0.68, 95 % CI 0.47-0.97) and for the ratio of serum betaine to choline (fourth vs first quartile adjusted OR 0.70, 95 % CI 0.48-1.00), but not for serum choline (fourth vs first quartile adjusted OR 0.80, 95 % CI 0.56-1.15). Serum betaine was inversely associated with breast cancer risk in subjects with below-median dietary folate intake (fourth vs first quartile adjusted OR 0.48, 95 % CI 0.30-0.77). CONCLUSIONS: This study suggested that serum betaine but not choline was inversely associated with breast cancer risk. This result needed to be further confirmed by the prospective studies.

Dietary choline and betaine intake, choline-metabolising genetic polymorphisms and breast cancer risk: a case-control study in China.

Choline and betaine are essential nutrients involved in one-carbon metabolism and have been hypothesised to affect breast cancer risk. Functional polymorphisms in genes encoding choline-related one-carbon metabolism enzymes, including phosphatidylethanolamine N-methyltransferase (PEMT), choline dehydrogenase (CHDH) and betaine-homocysteine methyltransferase (BHMT), have important roles in choline metabolism and may thus interact with dietary choline and betaine intake to modify breast cancer risk. This study aimed to investigate the interactive effect of polymorphisms in PEMT, BHMT and CHDH genes with choline/betaine intake on breast cancer risk among Chinese women. This hospital-based case-control study consecutively recruited 570 cases with histologically confirmed breast cancer and 576 age-matched (5-year interval) controls. Choline and betaine intakes were assessed by a validated FFQ, and genotyping was conducted for PEMT rs7946, CHDH rs9001 and BHMT rs3733890. OR and 95 % CI were estimated using unconditional logistic regression. Compared with the highest quartile of choline intake, the lowest intake quartile showed a significant increased risk of breast cancer. The SNP PEMT rs7946, CHDH rs9001 and BHMT rs3733890 had no overall association with breast cancer, but a significant risk reduction was observed among postmenopausal women with AA genotype of BHMT rs3733890 (OR 0·49; 95 % CI 0·25, 0·98). Significant interactions were observed between choline intake and SNP PEMT rs7946 (P interaction=0·029) and BHMT rs3733890 (P interaction=0·006) in relation to breast cancer risk. Our results suggest that SNP PEMT rs7946 and BHMT rs3733890 may interact with choline intake on breast cancer risk.

Joint effects of folate intake and one-carbon-metabolizing genetic polymorphisms on breast cancer risk: a case-control study in China.

This study aimed to examine the joint effects of folate intake, polymorphisms of 5,10- methylenetetrahydrofolate reductase (MTHFR), methionine synthesis reductase (MTRR) and methionine synthase (MTR) genes and breast cancer risk. A case-control study of 570 consecutively recruited breast cancer cases and 576 controls was conducted in Guangzhou, China. Multifactor dimensionality reduction and logistic regression approach were used to evaluate gene-gene interaction. The covariates were chosen based on comparison of baseline characteristics of cases and controls. Folate intake was found to be inversely associated with breast cancer risk. The MTRRrs162036 GG genotype was associated with a decreased risk of breast cancer [adjusted odds ratio (OR) 0.41, 95% confidence interval (CI) 0.20-0.85]. Compared with the wild-type group (MTRRrs162036 AA with MTRrs1805087 AA) MTRRrs162036 AA with MTRrs1805087 GA + GG was associated with a decreased risk (OR 0.70, 95% CI 0.48-1.03). With the combined MTHFRrs1801131 TT and MTHFRrs1801133 GG genotypes as a reference, MTHFRrs1801131 TT with MTHFRrs1801133 GA + AA was associated with a decreased risk (OR 0.78, 95% CI 0.57 - 1.08) and MTHFRrs1801131 GT + GG with MTHFRrs1801133 GA + AA was associated with an increased risk (OR 1.35, 95% CI 0.88-2.05). The joint impact of MTRRrs162036 and MTRrs1805087, MTHFRrs1801131 and MTHFRrs1801133, folate and MTHFRrs1801133 may contribute to breast cancer risk.

Specific serum carotenoids are inversely associated with breast cancer risk among Chinese women: a case-control study.

Previous epidemiological studies have revealed the anti-cancer effect of dietary circulating carotenoids. However, the protective role of specific individual circulating carotenoids has not been elucidated. The purpose of this study was to examine whether serum carotenoids, including α-carotene, ß-carotene, ß-cryptoxanthin, lycopene and lutein/zeaxanthin, could lower the risk for breast cancer among Chinese women. A total of 521 women with breast cancer and age-matched controls (5-year interval) were selected from three teaching hospitals in Guangzhou, China. Concentrations of α-carotene, ß-carotene, ß-cryptoxanthin, lycopene and lutein/zeaxanthin were measured using HPLC. Unconditional logistic regression models were used to calculate OR and 95% CI using quartiles defined in the control subjects. Significant inverse associations were observed between serum α-carotene, ß-carotene, lycopene, lutein/zeaxanthin and the risk for breast cancer. The multivariate OR for the highest quartile of serum concentration compared with the lowest quartile were 0·44 (95% CI 0·30, 0·65) for α-carotene, 0·27 (95% CI 0·18, 0·40) for ß-carotene, 0·41 (95% CI 0·28, 0·61) for lycopene and 0·26 (95% CI 0·17, 0·38) for lutein/zeaxanthin. However, no significant association was found between serum ß-cryptoxanthin and the risk for breast cancer. Stratified analysis by menopausal status and oestrogen receptor (ER)/progesterone receptor (PR) showed that serum α-carotene, ß-carotene, lycopene and lutein/zeaxanthin were inversely associated with breast cancer risk among premenopausal women and among all subtypes of ER or PR status. The results suggest a protective role of α-carotene, ß-carotene, lycopene and lutein/zeaxanthin, but not ß-cryptoxanthin, in breast cancer risk.

High serum iron level is associated with an increased risk of hypertensive disorders during pregnancy: a meta-analysis of observational studies.

The exact cause of hypertensive disorders in pregnancy (HDP) has not been clearly elucidated. Some researchers have recently investigated the relationship between the serum iron level and the incidence of HDP. However, the results are inconsistent, and these data have not been systematically evaluated. Therefore, we conducted a meta-analysis to evaluate the real association between the serum iron level and the incidence of HDP. We searched for published and ongoing trials in PubMed, EMBASE, Scopus, Web of Science, the Chinese Biomedical Database, CNKI, and the WANFANG database from January 1990 to May 2015 to identify studies that met our predefined criteria. Finally, 26 studies, including 1 cross-sectional study, 23 case-control studies, and 2 prospective nested case-control studies, including 1349 patients and 1119 control participants, were selected for this meta-analysis. The pooled results show that a high serum iron level increased the incidence of HDP (standard mean deviation [SMD], 1.50; 95% confidence interval [CI], 0.94-2.06; P < .0001), especially gestational hypertension (SMD, 3.65; 95% CI, 1.50-5.81; P = .0009) and preeclampsia (SMD, 1.27; 95% CI, 0.76-1.78; P < .0001). No significant difference was seen between the eclampsia groups and the control participants (SMD, 3.34; 95% CI, -0.02 to 6.69; P = .05). The results of this meta-analysis indicate that a high serum iron level is associated with an increased risk of HDP, especially gestational hypertension and preeclampsia.

High consumption of vegetable and fruit colour groups is inversely associated with the risk of colorectal cancer: a case-control study.

The colour of the edible portion of vegetables and fruit reflects the presence of specific micronutrients and phytochemicals. No existing studies have examined the relationship between the intake of vegetable and fruit colour groups and the risk of colorectal cancer. The present study, therefore, aimed to investigate these associations in a Chinese population. A case-control study was conducted between July 2010 and July 2014 in Guangzhou, China, in which 1057 consecutively recruited cases of colorectal cancer were frequency-matched to 1057 controls by age (5-year interval), sex and residence (rural/urban). A validated FFQ was used to collect dietary information during face-to-face interviews. Vegetables and fruit were classified into four groups according to the colour of their primarily edible parts: green; orange/yellow; red/purple; white. Unconditional logistic regression models were used to estimate the OR and 95 % CI. A higher consumption of orange/yellow, red/purple and white vegetables and fruit was inversely associated with the risk of colorectal cancer, with adjusted OR of 0·16 (95 % CI 0·12, 0·22) for orange/yellow, 0·23 (95 % CI 0·17, 0·31) for red/purple and 0·53 (95 % CI 0·40, 0·70) for white vegetables and fruit when the highest and lowest quartiles were compared. Total vegetable intake and total fruit intake have also been found to be inversely associated with colorectal cancer risk. However, the intake of green vegetable and fruit was not associated with colorectal cancer risk. The results of the present study, therefore, suggest that a greater intake of orange/yellow, red/purple and white vegetables and fruit is inversely associated with the risk of colorectal cancer.

Higher intake of carotenoid is associated with a lower risk of colorectal cancer in Chinese adults: a case-control study.

PURPOSE: The associations between specific carotenoid intake and colorectal cancer risk remain inconsistent. The aim of this study was to examine the association between specific dietary carotenoid intake with colorectal cancer risk in Chinese adults. METHOD: From July 2010 to October 2013, 845 eligible colorectal cancer cases and 845 frequency-matched controls (age and sex) completed in-person interviews. A validated food frequency questionnaire was used to estimate dietary intake. Multivariate logistical regression models were used to calculate the odds ratio (OR) and 95% confidence intervals (95% CIs) of colorectal cancer risk after adjusting for various confounders. RESULTS: A strong inverse association was found between ß-cryptoxanthin intake and colorectal cancer risk. Compared with the lowest quartile, the highest quartile intake showed a risk reduction of 77% (OR 0.23, 95% CI 0.17-0.33, P trend < 0.01) after adjustment for various confounding variables. The inverse associations were also observed for α-carotene (OR 0.50, 95% CI 0.37-0.68, P trend < 0.01), ß-carotene (OR 0.67, 95% CI 0.49-0.91, P trend < 0.01), and lycopene (OR 0.51, 95% CI 0.37-0.70, P trend < 0.01). There was no statistically significant association between lutein/zeaxanthin intake and colorectal cancer risk. These findings were consistent across cancer site, sources of controls, and smoking status. The inverse associations between dietary α-carotene, ß-cryptoxanthin, and lycopene intake and colorectal cancer risk were found in both males and females, while inverse associations between ß-carotene intake and colorectal cancer risk were only observed in males. CONCLUSIONS: Consumption of α-carotene, ß-carotene, ß-cryptoxanthin, and lycopene was inversely associated with colorectal cancer risk. No significant association was found between lutein/zeaxanthin intake and colorectal cancer risk.

SRC-3 is required for prostate cancer cell proliferation and survival.

Prostate cancer is the most common cancer in men in America. Currently, steroid receptor coactivators have been proposed to mediate the development and progression of prostate cancer, at times in a steroid-independent manner. Steroid receptor coactivator-3 (SRC-3, p/CIP, AIB1, ACTR, RAC3, and TRAM-1) is a member of the p160 family of coactivators for nuclear hormone receptors including the androgen receptor. SRC-3 is frequently amplified or overexpressed in a number of cancers. However, the role of SRC-3 in cancer cell proliferation and survival is still poorly understood. In this study, we show that SRC-3 is overexpressed in prostate cancer patients and its overexpression correlates with prostate cancer proliferation and is inversely correlated with apoptosis. Consistent with patient data, we have observed that reduction of SRC-3 expression by small interfering RNA decreases proliferation, delays the G1-S transition, and increases cell apoptosis of different prostate cancer cell lines. Furthermore, with decreased SRC-3 expression, proliferating cell nuclear antigen and Bcl-2 expression, as well as bromodeoxyuridine incorporation in prostate cancer cells are reduced. Finally, knockdown of SRC-3 with inducible short hairpin RNA expression in prostate cancer cells decreased tumor growth in nude mice. Taken together, these findings indicate that SRC-3 is an important regulator of prostate cancer proliferation and survival.

Inhibition of prostate tumor growth by overexpression of NudC, a microtubule motor-associated protein.

Microtubules play a central role in coordinating various cellular functions that are orchestrated by their interaction with molecular motors. Anticancer drugs that target microtubule dynamics have been shown to be effective in cancer treatment. However, the effect of microtubule motor-associated molecules on cancer cell proliferation is not clear. Here, we investigated the role of NudC, a nuclear movement protein associated with the microtubule motor dynein, on prostate tumorigenesis. Recombinant adenovirus expressing NudC (Ad-NudC) was used to examine the effects of NudC on the tumorigenicity of prostate cancer cells. Expression of NudC in LNCaP cells inhibited their anchorage-independent growth in a soft agar colony assay. Expression of NudC in DU145 or PC-3 cells inhibited tumor growth in a subcutaneous xenograft model. At the cellular level, expression of NudC in DU145 and PC-3 cells inhibited cell proliferation at 48 h after Ad-NudC infection. FACS analysis of cell cycle distribution showed that 50-60% of Ad-NudC-infected PC-3 cells have a G2/M-phase DNA content compared to about 16-19% in Ad-Luciferase (Ad-Luc)-infected control cells, suggesting that NudC overexpression resulted in aberrant cell cycle progression. Immunofluorescence microscopy revealed a significant increase in cells with a single enlarged nucleus and cells exhibiting multiple nuclei, along with a concomitant increase in cell size in Ad-NudC-infected cells. These results suggest that NudC overexpression led to a block in cell division of prostate cancer cells, and that Ad-NudC may provide a new anticancer drug approach targeting the function of a microtubule motor-associated protein.

Expression and purification of the angiogenesis inhibitor 16-kDa prolactin fragment from insect cells.

The 16-kDa fragment of prolactin (16-kDa PRL), derived from proteolytic cleavage of 23-kDa PRL, was shown to have antiangiogenic activity. Previous studies have shown that recombinant 16-kDa PRL produced from bacteria often contained endotoxins, which are cytotoxic to endothelial cells, and varied in its biological activity due to changes in its refolding from inclusion bodies. These problems limited the use of recombinant 16-kDa PRL. To improve the generation of recombinant 16-kDa PRL, we expressed 16-kDa PRL in Sf9 insect cells using a baculoviral expression system. The signal sequence of the human PRL gene and codons for seven histidines were added to the N- and C-termini, respectively, of the 16-kDa PRL cDNA construct. Recombinant 16-kDa PRL was detected in both the cell pellet and the medium. About 0.28 mg purified protein was isolated from the cell pellet of 4 x 10(7) infected cells using nickel affinity chromatography. Sixteen kilodalton PRL was posttranslationally modified with apparent molecular weights of 16 and 18 kDa on SDS-PAGE. The level of 18-kDa protein was significantly reduced after digestion with peptidyl-N-glycosidase, suggesting that the heterogeneity was due to glycosylation of 16-kDa PRL. N-terminal sequence analysis confirmed the fact that both proteins were human 16-kDa PRL and the signal sequences were cleaved at the same position as that of human PRL. Consistent with its role as an angiogenesis inhibitor, purified recombinant 16-kDa PRL inhibits the proliferation of endothelial cells with a potency similar to that previously reported for the protein generated in Escherichia coli. This 16-kDa PRL expressed in Sf9 cells is a useful reagent for functional studies and for the purification and identification of its receptor.

Role for NudC, a dynein-associated nuclear movement protein, in mitosis and cytokinesis.

NudC, a nuclear movement protein that associates with dynein, was originally cloned as a mitogen-inducible early growth response gene. NudC forms a biochemical complex with components of the dynein/dynactin complex and is suggested to play a role in translocation of nuclei in proliferating neuronal progenitors as well as in migrating neurons in culture. Here, we show that NudC plays multiple roles in mitosis and cytokinesis in cultured mammalian cells. Altering NudC levels by either small interfering RNA-mediated gene silencing or adenovirus-mediated overexpression resulted in multinucleated cells and cells with persistent intercellular connections and disorganized midzone and midbody matrix. These phenotypes suggest a failure in cytokinesis in NudC altered cells. Further, a key mitotic enzyme, polo-like kinase, is mislocalized from the centrosomes and the midbody in NudC altered cells. Gene silencing of nud-1, the Caenorhabditis elegans ortholog of NudC, led to a loss of midzone microtubules and the rapid regression of the cleavage furrow, which resulted in one-celled embryos containing two nuclei. The loss of midzone microtubule organization owing to silencing of the NudC/nud-1 gene in two systems, coupled with the loss of Plk1 from mitotic structures in mammalian cells, provide clues to the cytokinesis defect and the multinucleation phenotype. Our findings suggest that NudC functions in mitosis and cytokinesis, in part by regulating microtubule organization at the midzone and midbody.

Antitumor activity of the 16-kDa prolactin fragment in prostate cancer.

The 16-kDa prolactin (PRL), derived from the proteolytic cleavage of wild-type 23-kDa PRL, has been shown to have antiangiogenic activity. Such an antiangiogenic activity may have an effect on tumor growth in vivo. Here we examined the effect of 23-kDa and 16-kDa PRL on tumor growth, and the potential of using recombinant 16-kDa human PRL for prostate cancer therapy. The effects of 23-kDa PRL and 16-kDa PRL on the tumorigenicity of prostate cancer cells in vivo were studied. Using an adenovirus transfer vector to achieve high efficiency 23-kDa and 16-kDa PRL transfection in DU145 and PC-3 human prostate carcinoma cell lines, we demonstrated that expression of 16-kDa PRL in the prostate cancer cells markedly reduced their ability to form tumors in a xenograft animal model. These studies established that the 16-kDa PRL has antitumor activity in vivo, presumably as a result of its antiangiogenic effect. Interestingly, 23-kDa PRL showed a weak and transient suppression of prostate tumor growth. The weak antitumor activity of 23-kDa PRL may be because of the production of 16-kDa PRL from 23-kDa PRL by the tumor cells. Thus, the apparent effect of 23-kDa PRL on the growth of DU145 and PC-3 cells in vivo may result from the combined effects of 23-kDa PRL and 16-kDa PRL. These results suggest that the 16-kDa PRL has potential as a treatment agent in prostate cancer.

Hp95 promotes anoikis and inhibits tumorigenicity of HeLa cells.

p95 is a putative signal transduction protein of approximately 95 kDa that contains multiple tyrosine residues that are conserved from yeast to human, a Src phosphorylation consensus sequence and a proline-rich C-terminus that binds SH3-domains. Previous studies have established that mammalian p95 is physically associated with proteins that regulate apoptotic induction and cell transformation; however, it is unclear whether p95 is a positive or negative regulator in these processes. Moreover, a p95 partner protein has been localized at both focal adhesions and actin-cytoskeletons in rat astrocytes. However, there is no evidence that mammalian p95 has roles in regulating cell adhesion or morphology. In this study, we examined the effects of p95 on the anchorage-independent growth and tumorigenicity of malignant HeLa cells, and on the growth and morphology of non-transformed NIH3T3 cells. In HeLa cells, p95 overexpression promoted detachment-induced apoptosis (anoikis), inhibited detachment of viable cells from substratum and reduced tumorigenicity. In NIH3T3 cells, p95 overexpression promoted flat cell morphology and slowed cell proliferation, whereas p95 downregulation had opposite effects. These findings indicate that the mammalian p95 is a positive regulator in apoptotic signaling and a negative regulator in cell transformation. They also suggest that p95 has roles in regulating cell adhesion and morphology.

Enhanced suppression of prostate tumor growth by combining C-CAM1 gene therapy and angiogenesis inhibitor TNP-470.

We have previously shown that C-CAM1-based gene therapy effectively suppressed prostate tumor growth in nude mice xenograft models. In this study, we examined the effects of combining C-CAM1-based therapy and TNP-470, a potent angiogenesis inhibitor, on prostate cancer in a xenografted tumor model. The direct cytotoxic effects of Ad-C-CAM1 (recombinant adenovirus containing C-CAM1 cDNA) and TNP-470 on DU145 cells in vitro were determined by microculture tetrazolium assay. The in vivo antitumor effects of either agent alone were studied in a DU145 xenografted tumor model. Cells were infected with Ad-C-CAM1 or the control virus at multiplicities of infection (m.o.i.) of 5 or 10 and then inoculated onto nude mice 48 h later. TNP-470 (0, 17 or 35 mg/kg) was given 15, 17 and 19 days after inoculation. Combined treatments in vivo were carried out to determine whether there were synergistic antitumor effects. Both Ad-C-CAM1 and the control virus were minimally toxic to DU145 in vitro. There was evident dose-dependent suppression of xenografted tumor growth by either Ad-C-CAM1 or TNP-470. By the median-effect analysis, combination of the two agents generated strong synergistic antitumor effects as shown by marked tumor suppression as compared to either treatment alone. The novel strategy may have clinical implications for the treatment of prostate cancer.